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Hitman Jef Costello (Delon) lives in a single-room Parisian apartment whose spartan furnishings include a little bird in a cage. A long opening shot shows him lying on his bed, smoking, when the following text appears on-screen:

There is no greater solitude than that of the samurai unless it is that of the tiger in the jungle... Perhaps...

— Bushido (Book of the Samurai)

Costello's methodical modus operandi includes ironclad alibis involving his lover, Jane (Nathalie Delon). Having carried out a contract on a nightclub owner, he is seen leaving the scene by several witnesses, including piano player Valérie (Cathy Rosier). Their statements are inconsistent but the investigating officer (François Périer) believes Costello is his man. Costello loses a police tail and gets to a meeting point on a subway overpass where instead of getting paid, he is shot and wounded by a man sent by his employers. Having bandaged his wound and rested, he returns to the nightclub and somehow bonds with the piano player. In the meantime, police officers bug his room, which agitates the bird in its cage. Upon returning, Costello notices some loose feathers scattered around the cage. Suspecting an intrusion, he searches his room, finds the bug and deactivates it.

In the meantime, the police ransack Jane's apartment and offer her a deal: withdraw your alibi for Jef and we will leave you alone. Jane rejects the offer and shows them the door.

Back in his apartment Costello finds himself held at gunpoint by the overpass shooter who gives him money and offers him a new contract (the intended target is not revealed to the audience at this point). Costello overpowers him and forces him to disclose the identity of his boss, a man by the name of Olivier Rey (Jean-Pierre Posier).

Following a chase scene in the Métro and a goodbye visit to Jane, he drives to Rey's home, which turns out to be the same house in which the piano player lives. Costello kills Rey and drives to the nightclub.

This time he makes no attempt to conceal his presence. He even checks his hat but does not accept the hat-check ticket. Having put on his white gloves in full view of everyone, he walks over to the stage where Valérie advises him to leave. When he pulls out his gun and points it at her, she quietly asks "Why, Jef?" and he replies, "I was paid to." After a moment of staring, the audience hears gunshots, but not from his gun. Costello falls to the ground and dies. A junior police officer tells Valérie she is lucky the police were there because otherwise Costello would have killed her. But when his boss picks up Jef's gun, it is revealed that he had removed all the bullets before entering the club.

In an interview with Rui Nogueira, Melville indicated that he had shot an alternate version of Jef's death scene. In the alternative ending, which is actually the original version as Melville had written in the script, Costello meets his death with a picture-perfect grin à la Delon. The scene was changed to its current form when Melville angrily discovered that Delon had already used a smiling death scene in another of his films. Still images of the smiling death exist.

Hong Kong director John Woo's 1989 film, The Killer, was heavily influenced by Le Samouraï's plot, the bar's female pianist being replaced by a singer. Chow Yun-fat's character Jeffrey Chow (international character name for Ah Jong) was obviously inspired by Alain Delon's Jef Costello. The inspiration, or homage, is confirmed by the similarity in the character names. Woo acknowledged his influences by writing a short essay on Le Samouraï and Melville's techniques for the film's Criterion Collection DVD release.[4]

Hong Kong director Pang Ho-Cheung's 2001 crime-and-filmmaking comedy You Shoot, I Shoot features Eric Kot as a hitman who idolizes Alain Delon's Jef, dressing like the character, and speaking to him via a Le Samouraï poster in his apartment.[7]

Madonna's 2012 song "Beautiful Killer" is an homage to Alain Delon. The song alludes to Le Samouraï and Delon's Jef: "You are a beautiful killer / I like your silhouette when you stand on the streets / Like a samurai you can handle the heat / Makes me wanna pray for a haunted man..."[8]

The film is ranked #39 in Empire magazines "The 100 Best Films Of World Cinema" in 2010.[9]

The film is considered a cult classic and has the distinction of being one of the few films to have a 100% favorable rating at Rotten Tomatoes.[10]